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• W2997805557 abstract "HomePlant DiseaseVol. 104, No. 5First Report of Fusarium proliferatum Causing Leaf Sheath Rot on Clivia miniata in Henan Province, China PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Fusarium proliferatum Causing Leaf Sheath Rot on Clivia miniata in Henan Province, ChinaY. L. Li, Z. B. Yan, Y. H. Wang, and Z. ZhouY. L. Li†Corresponding author: Y. L. Li; E-mail Address: yonglili1978@163.comhttp://orcid.org/0000-0002-5615-1276College of Forestry, Henan University of Science and Technology, Luoyang, Henan 471023, ChinaSearch for more papers by this author, Z. B. YanCollege of Forestry, Henan University of Science and Technology, Luoyang, Henan 471023, ChinaSearch for more papers by this author, Y. H. WangCollege of Forestry, Henan University of Science and Technology, Luoyang, Henan 471023, ChinaSearch for more papers by this author, and Z. ZhouCollege of Forestry, Henan University of Science and Technology, Luoyang, Henan 471023, ChinaSearch for more papers by this authorAffiliationsAuthors and Affiliations Y. L. Li † Z. B. Yan Y. H. Wang Z. Zhou College of Forestry, Henan University of Science and Technology, Luoyang, Henan 471023, China Published Online:8 Mar 2020https://doi.org/10.1094/PDIS-12-18-2276-PDNAboutSectionsSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Clivia miniata (Lindl.) Verschaff is a popular indoor plant in China with attractive dark green foliage and vivid flowers. In summer 2016, leaf sheath rot (LSR) of pot-grown C. miniata was observed in a greenhouse and Anle ornamental market of Luoyang (34°39′N, 112°23′E), Henan Province. Symptoms were initial leaf sheath lesions, which turned into rot extending to one-third of the leaf, leading to death of the outer leaf. Tan sporodochia occurred on the lesions. Five plants were infected of 60 potted plants surveyed, and 25 leaves had LSR. Once a leaf exhibits LSR, the plant’s value significantly drops. Symptomatic leaves were cut into 1-cm2 pieces, surface sterilized in 70% ethanol for 30 s and 1% NaOCl for 5 min, and rinsed three times in sterilized distilled water. The disinfected symptomatic leaves were plated on PDA and incubated at 28°C for 5 days in darkness. Fusarium species were consistently isolated. Three isolates were obtained from single spores on different diseased plants and used for morphological and pathogenicity tests. Cultures on PDA produced white and floccose aerial mycelium, and light purple pigmentation was observed in the medium. The cottony hyphae of the pathogen aggregate into 2- to 5-mm-diameter clumps of mycelium in 2-week-old cultures on PDA. Macroconidia (n = 30) were sparse, slender, and mostly straight, with curved apical cell, 3 to 5 septate, 23.5 to 31.6 × 2.6 to 3.9 μm. Microconidia (n = 50) were abundant, single cell, oval or ellipsoid, hyaline, 2.9 to 11.0 × 1.5 to 2.6 μm. DNA was extracted from the three isolates/reisolates using a fungal DNA extraction kit (Sangon Biotech). The sequences of ITS, TUB2, and TEF1-α were amplified with PCR using primer pairs ITS1/ITS4 (White et al. 1990), Bt-2a/Bt-2b (Glass and Donaldson 1995), and ef1/ef2 (O’Donnell et al. 1998), respectively. The ITS, TUB2, and TEF1-α sequences are identical among the three isolates/reisolates. ITS (MF471668), a 558-bp amplicon, was 99% identical to MH055399.1 (Leyronas et al. 2018) and KJ128965 (Quesada-Ocampo et al. 2014). TUB2 (MG237917), a 337-bp amplicon, shared 99% identity with KJ128963 (Quesada-Ocampo et al. 2014) and AF291055 (Kwon et al. 2001). TEF1-α (MG237918), a 709-bp amplicon, shared 99% identity with KJ128964 (Quesada-Ocampo et al. 2014) and MH628463 (Leyronas et al. 2018). Multiple sequence alignments were generated using MAFFT v. 7 (Katoh and Standley 2013), and maximum parsimony analysis was performed on the multilocus combined TUB2 and TEF1-α alignment using PAPU v. 4.0b10. Clade stability of the trees was assessed by bootstrap analysis with 1,000 replicates. The isolate and strains of Fusarium proliferatum clustered into one clade (confidence level 100%). Species-specific primers CLPRO1/CLPRO2 (calmodulin gene) were used confirm identification (Mulè et al. 2004). A 526-bp band was observed in agarose gel from the isolates/reisolates. Based on morphological characteristics (Leslie and Summerell 2006), nucleotide analysis, and species-specific PCR, the fungus was identified as F. proliferatum. Surface-disinfested leaves of 2-year-old potted C. miniata plants were wounded with a sterile needle. Each isolate was inoculated on five plants. The two lateral leaves were inoculated with 5 μl of 2 × 107 conidia/ml suspension on each wound. Each isolate was inoculated on 10 lower leaves (one isolate per leaf). Ten leaves on 2-year-old plants were inoculated with sterile water. Plants were incubated at 28°C and 85% RH. All leaves inoculated with F. proliferatum developed faint yellow lesions after 2 days, which enlarged over time; control plants were symptomless. F. proliferatum was reisolated from symptomatic leaves, confirming Koch’s postulates. F. proliferatum causes many important crop diseases, including crown rot of asparagus, wilt disease on carnation and sunflower, mango malformation disease, and leaf spot on tobacco (Li et al. 2017). This is the first report of F. proliferatum causing LSR on C. miniata in China.The author(s) declare no conflict of interest.References:Glass, N. L., and Donaldson, G. C. 1995. Appl. Environ. Microbiol. 61:1323. Crossref, ISI, Google ScholarKatoh, K., and Standley, D. M. 2013. Mol. Biol. Evol. 30:772. https://doi.org/10.1093/molbev/mst010 Crossref, ISI, Google ScholarKwon, S. I., et al. 2001. Can. J. Bot. 79:1115. Crossref, Google ScholarLeslie, J. F., and Summerell, B. A., eds. 2006. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA. doi.org/10.1002/9780470278376 Crossref, Google ScholarLeyronas, S. C., et al. 2018. Plant Dis. 102:2658. https://doi.org/10.1094/PDIS-06-18-0962-PDN Link, ISI, Google ScholarLi, F. J., et al. 2017. J. Gen. Plant Pathol. 83:264. https://doi.org/10.1007/s10327-017-0714-y Crossref, ISI, Google ScholarMulè, G., et al. 2004. FEMS Microbiol. Lett. 230:235. https://doi.org/10.1016/S0378-1097(03)00926-1 Crossref, ISI, Google ScholarO’Donnell, K., et al. 1998. Proc. Natl. Acad. Sci. U.S.A. 95:2044. https://doi.org/10.1073/pnas.95.5.2044 Crossref, ISI, Google ScholarQuesada-Ocampo, L. M., et al. 2014. Plant Dis. 98:1009. https://doi.org/10.1094/PDIS-01-14-0040-PDN Link, ISI, Google ScholarWhite, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA. Crossref, Google ScholarThe author(s) declare no conflict of interest.Funding: Funding was provided by National Natural Science Foundation of China (31600519).DetailsFiguresLiterature CitedRelated Vol. 104, No. 5 May 2020SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionSymptoms observed in the field on zucchini plants caused by Fusarium solani f. sp. cucurbitae (A. Pérez-Hernández et al.). Photo credit: J. M. Gómez-Vázquez. Peach tree with excavated root collar (S. B. Miller et al.). Photo credit: G. Schnabel. Metrics Article History Issue Date: 3 May 2020Published: 8 Mar 2020First Look: 26 Dec 2019Accepted: 23 Dec 2019 Page: 1552 Information© 2020 The American Phytopathological SocietyFundingNational Natural Science Foundation of ChinaGrant/Award Number: 31600519Keywordsfungipathogen detectionornamentalsherbaceous/flowering plantsThe author(s) declare no conflict of interest.Cited byIdentification, Pathogenicity, and Genetic Diversity of Fusarium spp. Associated with Maize Sheath Rot in Heilongjiang Province, China16 September 2022 | International Journal of Molecular Sciences, Vol. 23, No. 18Fusarium proliferatumCABI Compendium, Vol. CABI CompendiumMicromorphology and Anatomy of the Flowers in Clivia spp. and Scadoxus multiflorus (Haemantheae, Amaryllidaceae)9 December 2021 | Acta Agrobotanica, Vol. 74Морфологія, фенологія та плодоношення Clivia miniata (Lindl.) Verschaff. (Amaryllidaceae J.St.-Hil.) у культурі15 September 2020 | Lesya Ukrainka Eastern European National University Scientific Bulletin. Series: Biological Sciences, No. 1(389)" @default.
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